Determination of genetic diversity of Leishmania species using mini-circle kDNA, in Iran-Iraq countries border.

Department of Medical Parasitology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

Abstract

Background and Objective:

Cutaneous leishmaniasis (CL) is one of the most important diseases worldwide, with a different range of prevalence in endemic areas. Anthroponotic and zoonotic CL are two epidemiological forms of CL, in Iran. Although Ilam Province in the west of Iran is one of the main endemic areas of the disease, there is no inclusive study to determine the genetic variations of parasite in these areas. The objective of this study was to determine the genetic diversity of Leishmania species in Ilam Province, using mini-circle kDNA gene.

Materials and Methods:

Direct smears were taken from skin lesions of 200 suspected cases of CL. Smears were stained, screened under light microscope. Polymerase chain reaction (PCR) was performed, using specific kinetoplast DNA primers. Data were analyzed, using the molecular bio-software.

Results:

All the samples were positive by direct examination. PCR results showed all cases were positive for Leishmania major. Although all isolated cases belong to a different county of Ilam province, all were positive for L. major with intra-species genetic diversity, divided into four clades in the dendrogram.

Interpretation and Conclusion:

This variation can affect drug resistance and controlling strategies of parasite. It is possible that different species of sand flies and rodents are the vector and reservoir of parasite, respectively; however, further studies are needed to validate this.

Phylogenetic tree of the kDNA gene sequences of Leishmania isolates constructed by the neighbor-joining method using Mega6 software (version 6). Isolates or strains names are as provided in Genbank as available. Sequences generated in this study named as Leishmania major IR/Il. Leishmania tropica (accession number AB901375.1) used as the out-group